Consortia for High-Throughput-Enabled Structural Biology Partnerships (U01)

高通量结构生物学合作联盟 (U01)

• 批准号: 8149802
• 负责人: Joshua N. Adkins
• 金额: $ 100.75万
• 依托单位: BATTELLE PACIFIC NORTHWEST LABORATORIES
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-30 至 2015-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8149802
• 关键词: Consortia; Throughput; Enabled; Structural; Biology

DESCRIPTION (provided by applicant): While powerful for studying pathogenic secreted effectors, structural biology has not been used to a large extent to study effector-host protein interactions. In this proposal, we use known and novel secreted effectors and an experimental pipeline that will provide broad insights into how a model pathogen, Salmonella, subverts host cell function. Aim 1 simultaneously provides key functional insights into effector protein potency and function using broad set of assays, while at the same time serves as an important selection step to focus structural characterization by the PSI network on the most valuable targets. Aim 2 provides valuable information about the host proteins and protein pathways, using cross-linking and proteomics, that each effector interacts with and provides the PSI network with a prioritized list of host protein targets for structural characterization. Aim 3 interrogates possible specific protein-protein interactions and provides a selection of validated protein-protein interactions and possibly ligands for structural characterization by PSI network. Ultimately, structure-function studies of individual secreted effectors are invaluable to the host-pathogen research community. However, the insights into host-pathogen biology gained from this large parallel characterization effort with multiple effectors will advance understanding at a more complete systems level. PUBLIC HEALTH RELEVANCE: Advanced mass spectrometry methods with structure determinations will allow for improved understanding of the interactions between human host proteins and pathogens; this research may lead to new therapies for infectious diseases. The specific pathogen model to be studied here is Salmonella, a pathogen that has been the cause of a number of significant food-borne outbreaks in recent years.

描述（由申请人提供）：虽然结构生物学对于研究致病性分泌效应子很有效，但尚未在很大程度上用于研究效应子-宿主蛋白相互作用。在这项提案中，我们使用已知的和新颖的分泌效应器和实验管道，这将为模型病原体沙门氏菌如何破坏宿主细胞功能提供广泛的见解。目标 1 同时使用广泛的检测方法提供对效应蛋白效力和功能的关键功能见解，同时作为重要的选择步骤，将 PSI 网络的结构表征集中在最有价值的靶标上。 目标 2 使用交联和蛋白质组学提供有关宿主蛋白质和蛋白质途径的有价值的信息，每个效应器与宿主蛋白质和蛋白质途径相互作用，并为 PSI 网络提供宿主蛋白质目标的优先列表，以进行结构表征。目标 3 询问可能的特定蛋白质-蛋白质相互作用，并提供一系列经过验证的蛋白质-蛋白质相互作用和可能的配体，用于 PSI 网络的结构表征。最终，个体分泌效应器的结构功能研究对于宿主-病原体研究界来说是无价的。然而，从这种具有多个效应器的大型并行表征工作中获得的对宿主-病原体生物学的见解将在更完整的系统水平上促进理解。 公共卫生相关性：具有结构测定的先进质谱方法将有助于更好地了解人类宿主蛋白和病原体之间的相互作用；这项研究可能会带来传染病的新疗法。这里要研究的特定病原体模型是沙门氏菌，这种病原体是近年来许多重大食源性疫情爆发的原因。